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Sélection d’aptamères anti-adénine ADN modifiés en présence de solvant organique. Application au développement de biocapteurs

Abstract : Development of in situ detection tools (IDT) is required for specific real time monitoringof chemical species. Efficient IDT is not only related to its affinity and ability todiscriminate between molecular variants, but moreover it must be adapted for operatingunder conditions imposed by the context of application. Among others, hightemperature, pH variation and presence of organic solvents may be mentioned. The aimof our project is to develop an aptamer operating in the presence of organic solvents. Forthis purpose, we opted for selection in presence of methanol. Limitations of this strategyare adaptability of SELEX technology and chemical diversity of nucleic acids. For thispurpose, we used library incorporating (5-(octa1,7-diynyl)-2’-deoxyuridine) dDOTPinstead of conventional thymine nucleotide. Our experimental strategy led to theselection of an aptamer that specifically recognises adenine in the presence of 25 % ofmethanol. dDOTP nucleotides are essential for adenine recognition; the selectedaptamer shares a purine rich motif with a previously described adenosine aptamer, butdisplays a specific structure motif, essential for operating in the presence of methanol.Furthermore, the ability of truncated variants of the selected aptamer to form arecognition module of biosensor was assessed in this study.
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Thinhinane Chaou. Sélection d’aptamères anti-adénine ADN modifiés en présence de solvant organique. Application au développement de biocapteurs. Autre [q-bio.OT]. Université de Bordeaux, 2015. Français. ⟨NNT : 2015BORD0385⟩. ⟨tel-01674234⟩

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